The composition of microbial communities and the functioning of ecosystems are greatly influenced by the nutrient inputs. Despite this fact, our knowledge regarding the impact of phosphorus (P) inputs on soil P availability and microbial community structures in subtropical acidic soils remains limited. We hypothesized that diverse P inputs, incubation temperatures, and soil types could significantly alter soil P availability and microbial communities. To address this gap, we conducted a laboratory incubation experiment, investigating the effects of biochar and inorganic P amendments on soil available P, soil pH, acid phosphatase enzymes, and bacterial abundance. We employed two different incubation temperatures (15 °C and 25 °C) using acidic paddy soil and red soil from the subtropical Southern China region. Our results indicate a notable increase in soil pH, reaching 37% and 39% at 15 °C and 40% and 40.6% at 25 °C, respectively, following the application of biochar and inorganic P amendments in paddy soil. In the case of red soil, we observed pH increases of 41% and 43% at 15 °C and 44% and 45% at 25 °C after the application of biochar and inorganic P amendment, respectively. The addition of inorganic P amendment resulted in the highest available P contents in paddy soil, reaching 111.47 mg/kg at 15 °C and 100.17 mg/kg at 25 °C, respectively. However, Proteobacteria decreased after inorganic P addition, which showed that P might not be the only limiting nutrient for various bacterial communities. Bacterial diversity and richness indices were found to be higher after biochar application in both soils. Gemmatimonadetes, Acidobacteria, and Actinobacteria were found to be strongly influenced by incubation temperatures, whereas most of the top abundant bacterial phyla, such as Gemmatimonadetes, Actinobacteria, Chloroflexi, Acidobacteria, Planctomycetes, Firmicutes, Patescibacteria, and Bacteroidetes, were highly dependent on soil type. At the genus level, various important P solubilizing genera (Pseudomonas, Bradyrhizobium, Streptomyces jietaisiensis, Massilia) significantly increased after biochar and inorganic P addition in both soils. The correlation analysis proved that P-solubilizing genera were significantly associated with changes in soil pH, as well as soil available P after biochar and inorganic P addition. Conclusively, in a short-term incubation experiment, inorganic P amendment greatly increased the soil pH and available phosphorus contents compared to biochar and control treatments; however, the microbial community was observed to be strongly associated with biochar application, soil type, and incubation temperature.